Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Journal of Experimental & Clinical Cancer Research
Published in: Journal of Experimental & Clinical Cancer Research 1/2024

Open Access 01-12-2024 | Glioma | Research

Functionally-instructed modifiers of response to ATR inhibition in experimental glioma

Authors: Bianca Walter, Sophie Hirsch, Laurence Kuhlburger, Aaron Stahl, Leonard Schnabel, Silas Wisser, Lara A. Haeusser, Foteini Tsiami, Sarah Plöger, Narges Aghaallaei, Advaita M Dick, Julia Skokowa, Christian Schmees, Markus Templin, Katja Schenke-Layland, Marcos Tatagiba, Sven Nahnsen, Daniel J. Merk, Ghazaleh Tabatabai

Published in: Journal of Experimental & Clinical Cancer Research | Issue 1/2024

Login to get access

Abstract

Background

The DNA damage response (DDR) is a physiological network preventing malignant transformation, e.g. by halting cell cycle progression upon DNA damage detection and promoting DNA repair. Glioblastoma are incurable primary tumors of the nervous system and DDR dysregulation contributes to acquired treatment resistance. Therefore, DDR targeting is a promising therapeutic anti-glioma strategy. Here, we investigated Ataxia telangiectasia and Rad3 related (ATR) inhibition (ATRi) and functionally-instructed combination therapies involving ATRi in experimental glioma.

Methods

We used acute cytotoxicity to identify treatment efficacy as well as RNAseq and DigiWest protein profiling to characterize ATRi-induced modulations within the molecular network in glioma cells. Genome-wide CRISPR/Cas9 functional genomic screens and subsequent validation with functionally-instructed compounds and selected shRNA-based silencing were employed to discover and investigate molecular targets modifying response to ATRi in glioma cell lines in vitro, in primary cultures ex vivo and in zebrafish and murine models in vivo.

Results

ATRi monotherapy displays anti-glioma efficacy in vitro and ex vivo and modulates the molecular network. We discovered molecular targets by genome-wide CRISPR/Cas9 loss-of-function and activation screens that enhance therapeutic ATRi effects. We validated selected druggable targets by a customized drug library and functional assays in vitro, ex vivo and in vivo.

Conclusion

In conclusion, our study leads to the identification of novel combination therapies involving ATRi that could inform future preclinical studies and early phase clinical trials.
Appendix
Available only for authorised users
Literature
1.
2.
Pilié PG, Tang C, Mills GB, Yap TA. State-of-the-art strategies for targeting the DNA damage response in cancer. Nat Reviews Clin Oncol. 2019;16(2):81–104.CrossRef
3.
Pearl LH, Schierz AC, Ward SE, Al-Lazikani B, Pearl FMG. Therapeutic opportunities within the DNA damage response. Nat Rev Cancer. 2015;15(3):166–80.PubMedCrossRef
4.
Roos WP, Thomas AD, Kaina B. DNA damage and the balance between survival and death in cancer biology. Nat Rev Cancer. 2016;16(1):20–33.PubMedCrossRef
5.
Taylor AMR, Rothblum-Oviatt C, Ellis NA, Hickson ID, Meyer S, Crawford TO, et al. Chromosome instability syndromes. Nat Reviews Disease Primers. 2019;5(1):64.PubMedCrossRef
6.
7.
Hanahan D, Weinberg Robert A. Hallmarks of Cancer: the Next Generation. Cell. 2011;144(5):646–74.PubMedCrossRef
8.
Bryant HE, Schultz N, Thomas HD, Parker KM, Flower D, Lopez E, et al. Specific killing of BRCA2-deficient tumours with inhibitors of poly(ADP-ribose) polymerase. Nature. 2005;434(7035):913–7.ADSPubMedCrossRef
9.
Fong PC, Boss DS, Yap TA, Tutt A, Wu P, Mergui-Roelvink M, et al. Inhibition of poly(ADP-Ribose) polymerase in tumors from BRCA mutation carriers. N Engl J Med. 2009;361(2):123–34.PubMedCrossRef
10.
Groelly FJ, Fawkes M, Dagg RA, Blackford AN, Tarsounas M. Targeting DNA damage response pathways in cancer. Nat Rev Cancer. 2023;23(2):78–94.PubMedCrossRef
11.
Weller M, van den Bent M, Preusser M, Le Rhun E, Tonn JC, Minniti G, et al. EANO guidelines on the diagnosis and treatment of diffuse gliomas of adulthood. Nat Reviews Clin Oncol. 2021;18(3):170–86.CrossRef
12.
Stupp R, Taillibert S, Kanner A, Read W, Steinberg DM, Lhermitte B, et al. Effect of Tumor-Treating Fields Plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: a Randomized Clinical Trial. JAMA. 2017;318(23):2306–16.PubMedPubMedCentralCrossRef
13.
Stupp R, Hegi ME, Mason WP, van den Bent MJ, Taphoorn MJB, Janzer RC, et al. Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC-NCIC trial. Lancet Oncol. 2009;10(5):459–66.PubMedCrossRef
14.
Stupp R, Mason WP, van den Bent MJ, Weller M, Fisher B, Taphoorn MJB, et al. Radiotherapy plus Concomitant and Adjuvant Temozolomide for Glioblastoma. N Engl J Med. 2005;352(10):987–96.PubMedCrossRef
15.
16.
Bao S, Wu Q, McLendon RE, Hao Y, Shi Q, Hjelmeland AB, et al. Glioma stem cells promote radioresistance by preferential activation of the DNA damage response. Nature. 2006;444(7120):756–60.ADSPubMedCrossRef
17.
Carruthers RD, Ahmed SU, Ramachandran S, Strathdee K, Kurian KM, Hedley A, et al. Replication stress drives constitutive activation of the DNA damage response and Radioresistance in Glioblastoma Stem-like cells. Cancer Res. 2018;78(17):5060–71.PubMedPubMedCentralCrossRef
18.
Sundar R, Brown J, Ingles Russo A, Yap TA. Targeting ATR in cancer medicine. Curr Probl Cancer. 2017;41(4):302–15.PubMedCrossRef
19.
Dillon MT, Espinasse A, Ellis S, Mohammed K, Grove LG, McLellan L, et al. Abstract CT084: a phase I dose-escalation study of ATR inhibitor monotherapy with AZD6738 in advanced solid tumors (PATRIOT part A). Cancer Res. 2017;77(13Supplement):CT084–CT.CrossRef
20.
Koch MS, Czemmel S, Lennartz F, Beyeler S, Rajaraman S, Przystal JM et al. Experimental glioma with high bHLH expression harbor increased replicative stress and are sensitive toward ATR inhibition. Neuro-Oncology Adv. 2020;2(1).
21.
Jackson CB, Noorbakhsh SI, Sundaram RK, Kalathil AN, Ganesa S, Jia L, et al. Temozolomide sensitizes MGMT-Deficient tumor cells to ATR inhibitors. Cancer Res. 2019;79(17):4331–8.PubMedPubMedCentralCrossRef
22.
Koch MS, Zdioruk M, Nowicki MO, Griffith AM, Aguilar-Cordova E, Aguilar LK, et al. Perturbing DDR signaling enhances cytotoxic effects of local oncolytic virotherapy and modulates the immune environment in glioma. Mol Therapy - Oncolytics. 2022;26:275–88.CrossRef
23.
Yap TA, Krebs MG, Postel-Vinay S, El-Khouiery A, Soria J-C, Lopez J, et al. Ceralasertib (AZD6738), an oral ATR Kinase Inhibitor, in combination with carboplatin in patients with Advanced Solid tumors: a phase I study. Clin Cancer Res. 2021;27(19):5213–24.PubMedPubMedCentralCrossRef
24.
Fròsina G, Profumo A, Marubbi D, Marcello D, Ravetti JL, Daga A. ATR kinase inhibitors NVP-BEZ235 and AZD6738 effectively penetrate the brain after systemic administration. Radiat Oncol. 2018;13(1):76.PubMedPubMedCentralCrossRef
25.
Talele S, Zhang W, Burgenske DM, Kim M, Mohammad AS, Dragojevic S, et al. Brain distribution of Berzosertib: an Ataxia Telangiectasia and Rad3-Related protein inhibitor for the treatment of Glioblastoma. J Pharmacol Exp Ther. 2021;379(3):343–57.PubMedPubMedCentralCrossRef
26.
Treindl F, Ruprecht B, Beiter Y, Schultz S, Döttinger A, Staebler A, et al. A bead-based western for high-throughput cellular signal transduction analyses. Nat Commun. 2016;7(1):12852.ADSPubMedPubMedCentralCrossRef
27.
Ishii N, Maier D, Merlo A, Tada M, Sawamura Y, Diserens A-C, Van Meir EG. Frequent co-alterations of TP53, p16/CDKN2A, p14ARF, PTEN Tumor suppressor genes in human glioma cell lines. Brain Pathol. 1999;9(3):469–79.PubMedCrossRef
28.
Kanehisa M, Furumichi M, Sato Y, Ishiguro-Watanabe M, Tanabe M. KEGG: integrating viruses and cellular organisms. Nucleic Acids Res. 2020;49(D1):D545–D51.PubMedCentralCrossRef
29.
Bruce JL, Hurford RK, Classon M, Koh J, Dyson N. Requirements for cell cycle arrest by p16INK4a. Mol Cell. 2000;6(3):737–42.PubMedCrossRef
30.
Doench JG, Fusi N, Sullender M, Hegde M, Vaimberg EW, Donovan KF, et al. Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9. Nat Biotechnol. 2016;34(2):184–91.PubMedPubMedCentralCrossRef
31.
Sanson KR, Hanna RE, Hegde M, Donovan KF, Strand C, Sullender ME, et al. Optimized libraries for CRISPR-Cas9 genetic screens with multiple modalities. Nat Commun. 2018;9(1):5416.ADSPubMedPubMedCentralCrossRef
32.
Li W, Zhang H, Assaraf YG, Zhao K, Xu X, Xie J, et al. Overcoming ABC transporter-mediated multidrug resistance: molecular mechanisms and novel therapeutic drug strategies. Drug Resist Updates. 2016;27:14–29.CrossRef
33.
Farmer H, McCabe N, Lord CJ, Tutt ANJ, Johnson DA, Richardson TB, et al. Targeting the DNA repair defect in BRCA mutant cells as a therapeutic strategy. Nature. 2005;434(7035):917–21.ADSPubMedCrossRef
34.
Kaufman B, Shapira-Frommer R, Schmutzler RK, Audeh MW, Friedlander M, Balmaña J, et al. Olaparib Monotherapy in patients with Advanced Cancer and a germline BRCA1/2 mutation. J Clin Oncol. 2015;33(3):244–50.PubMedCrossRef
35.
Herrlinger U, Tzaridis T, Mack F, Steinbach JP, Schlegel U, Sabel M, et al. Lomustine-temozolomide combination therapy versus standard temozolomide therapy in patients with newly diagnosed glioblastoma with methylated MGMT promoter (CeTeG/NOA–09): a randomised, open-label, phase 3 trial. Lancet. 2019;393(10172):678–88.PubMedCrossRef
36.
Zhang J, Stevens FG, Bradshaw MD, Temozolomide T. Mechanisms of action, repair and resistance. Curr Mol Pharmacol. 2012;5(1):102–14.PubMedCrossRef
37.
Mojas N, Lopes M, Jiricny J. Mismatch repair-dependent processing of methylation damage gives rise to persistent single-stranded gaps in newly replicated DNA. Genes Dev. 2007;21(24):3342–55.PubMedPubMedCentralCrossRef
38.
Derby S, Dutton L, Strathdee KE, Stevenson K, Koessinger A, Jackson M et al. Inhibition of ATR opposes glioblastoma invasion through disruption of cytoskeletal networks and integrin internalisation via macropinocytosis. Neurooncology. 2023.
39.
40.
41.
Günther HS, Schmidt NO, Phillips HS, Kemming D, Kharbanda S, Soriano R, et al. Glioblastoma-derived stem cell-enriched cultures form distinct subgroups according to molecular and phenotypic criteria. Oncogene. 2008;27(20):2897–909.PubMedCrossRef
42.
Serra V, Wang AT, Castroviejo-Bermejo M, Polanska UM, Palafox M, Herencia-Ropero A, et al. Identification of a molecularly-defined subset of breast and ovarian Cancer models that respond to WEE1 or ATR Inhibition, overcoming PARP inhibitor resistance. Clin Cancer Res. 2022;28(20):4536–50.PubMedPubMedCentralCrossRef
43.
Kitajima M, Iwai M, Kikura-Hanajiri R, Goda Y, Iida M, Yabushita H, Takayama H. Discovery of indole alkaloids with cannabinoid CB1 receptor antagonistic activity. Bioorg Med Chem Lett. 2011;21(7):1962–4.PubMedCrossRef
44.
Meschini S, Marra M, Condello M, Calcabrini A, Federici E, Dupuis ML, et al. Voacamine, an alkaloid extracted from Peschiera Fuchsiaefolia, inhibits P-glycoprotein action in multidrug-resistant tumor cells. Int J Oncol. 2005;27(6):1597–603.PubMed
45.
Tentori L, Ricci-Vitiani L, Muzi A, Ciccarone F, Pelacchi F, Calabrese R, et al. Pharmacological inhibition of poly(ADP-ribose) polymerase-1 modulates resistance of human glioblastoma stem cells to temozolomide. BMC Cancer. 2014;14(1):151.PubMedPubMedCentralCrossRef
46.
Leijen S, Geel RMJMv, Sonke GS, Jong Dd, Rosenberg EH, Marchetti S, et al. Phase II study of WEE1 inhibitor AZD1775 plus carboplatin in patients with TP53-Mutated ovarian Cancer Refractory or resistant to First-Line Therapy within 3 months. J Clin Oncol. 2016;34(36):4354–61.PubMedCrossRef
47.
Chapman T, Kinsella T. Ribonucleotide reductase inhibitors: a New look at an old target for Radiosensitization. Front Oncol. 2012;1.
48.
Krönke J, Narla A, Hurst SN, Udeshi N, Schenone M, McConkey M, et al. Inhibition of the CRBN-DDB1-CUL4-ROC1 E3 ubiquitin ligase mediates the anti-proliferative and Immunomodulatory properties of Lenalidomide. Blood. 2012;120(21):919.CrossRef
49.
Wilson Z, Odedra R, Wallez Y, Wijnhoven PWG, Hughes AM, Gerrard J, et al. ATR inhibitor AZD6738 (Ceralasertib) exerts Antitumor Activity as a Monotherapy and in combination with chemotherapy and the PARP inhibitor Olaparib. Cancer Res. 2022;82(6):1140–52.PubMedPubMedCentralCrossRef
50.
Chen T, Yang F, Dai X, Yu Y, Sun Y, Wu X, et al. VE-822 enhanced Cisplatin Chemotherapy effects on Head and Neck squamous cell Carcinoma Drug-resistant cells. Curr Cancer Drug Targets. 2023;23(6):482–95.PubMedCrossRef
51.
Shapiro GI, Wesolowski R, Devoe C, Lord S, Pollard J, Hendriks BS, et al. Phase 1 study of the ATR inhibitor berzosertib in combination with cisplatin in patients with advanced solid tumours. Br J Cancer. 2021;125(4):520–7.PubMedPubMedCentralCrossRef
52.
Blackford AN, Jackson SP, ATM, ATR. The trinity at the heart of the DNA damage response. Mol Cell. 2017;66(6):801–17.PubMedCrossRef
53.
Carminati PO, Donaires FS, Marques MM, Donadi EA, Passos GAS, Sakamoto-Hojo ET. Cisplatin associated with LY294002 increases cytotoxicity and induces changes in transcript profiles of glioblastoma cells. Mol Biol Rep. 2014;41(1):165–77.PubMedCrossRef
54.
Walter B, Canjuga D, Yüz SG, Ghosh M, Bozko P, Przystal JM, et al. Argyrin F Treatment-Induced vulnerabilities lead to a Novel Combination Therapy in Experimental Glioma. Adv Ther. 2021;4(9):2100078.CrossRef
55.
Anderle N, Koch A, Gierke B, Keller A-L, Staebler A, Hartkopf A, et al. A platform of patient-derived Microtumors identifies individual treatment responses and therapeutic vulnerabilities in Ovarian Cancer. Cancers. 2022;14(12):2895.PubMedPubMedCentralCrossRef
56.
Przystal JM, Becker H, Canjuga D, Tsiami F, Anderle N, Keller A-L, et al. Targeting CSF1R alone or in combination with PD1 in experimental glioma. Cancers. 2021;13(10):2400.PubMedPubMedCentralCrossRef
57.
Guzmán C, Bagga M, Kaur A, Westermarck J, Abankwa D. ColonyArea: an ImageJ Plugin to automatically quantify colony formation in Clonogenic assays. PLoS ONE. 2014;9(3):e92444.ADSPubMedPubMedCentralCrossRef
58.
59.
60.
Raudvere U, Kolberg L, Kuzmin I, Arak T, Adler P, Peterson H, Vilo J. G:profiler: a web server for functional enrichment analysis and conversions of gene lists (2019 update). Nucleic Acids Res. 2019;47(W1):W191–W8.PubMedPubMedCentralCrossRef
61.
Saeed AI, Sharov V, White J, Li J, Liang W, Bhagabati N, et al. TM4: a free, open-source system for Microarray Data Management and Analysis. Biotechniques. 2003;34(2):374–8.PubMedCrossRef
62.
63.
Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature. 2015;517(7536):583–8.ADSPubMedCrossRef
64.
Li W, Köster J, Xu H, Chen C-H, Xiao T, Liu JS, et al. Quality control, modeling, and visualization of CRISPR screens with MAGeCK-VISPR. Genome Biol. 2015;16(1):281.PubMedPubMedCentralCrossRef
65.
Yan H, Zhang B, Li S, Zhao Q. A formal model for analyzing drug combination effects and its application in TNF-α-induced NFκB pathway. BMC Syst Biol. 2010;4(1):50.PubMedPubMedCentralCrossRef
66.
He L, Kulesskiy E, Saarela J, Turunen L, Wennerberg K, Aittokallio T, Tang J. Methods for high-throughput drug combination screening and synergy scoring. In: von Stechow L, editor. Cancer systems Biology: methods and protocols. New York, NY: Springer New York; 2018. pp. 351–98.CrossRef
67.
68.
Sancak Y, Peterson TR, Shaul YD, Lindquist RA, Thoreen CC, Bar-Peled L, Sabatini DM. The rag GTPases bind Raptor and mediate amino acid signaling to mTORC1. Science. 2008;320(5882):1496–501.ADSPubMedPubMedCentralCrossRef
Metadata
Title
Functionally-instructed modifiers of response to ATR inhibition in experimental glioma
Authors
Bianca Walter
Sophie Hirsch
Laurence Kuhlburger
Aaron Stahl
Leonard Schnabel
Silas Wisser
Lara A. Haeusser
Foteini Tsiami
Sarah Plöger
Narges Aghaallaei
Advaita M Dick
Julia Skokowa
Christian Schmees
Markus Templin
Katja Schenke-Layland
Marcos Tatagiba
Sven Nahnsen
Daniel J. Merk
Ghazaleh Tabatabai
Publication date
01-12-2024
Publisher
BioMed Central
Published in
Journal of Experimental & Clinical Cancer Research / Issue 1/2024
Electronic ISSN: 1756-9966
DOI
https://doi.org/10.1186/s13046-024-02995-z

Other articles of this Issue 1/2024

Journal of Experimental & Clinical Cancer Research 1/2024 Go to the issue

Research

PRMT6-mediated transcriptional activation of ythdf2 promotes glioblastoma migration, invasion, and emt via the wnt–β-catenin pathway

Research

PSMD9 promotes the malignant progression of hepatocellular carcinoma by interacting with c-Cbl to activate EGFR signaling and recycling

Review

Mitochondrial transfer in tunneling nanotubes—a new target for cancer therapy

Research

LOXL1-AS1 contributes to metastasis in sonic-hedgehog medulloblastoma by promoting cancer stem-like phenotypes

Research

TRIM25 promotes glioblastoma cell growth and invasion via regulation of the PRMT1/c-MYC pathway by targeting the splicing factor NONO

Research

PAX6 promotes neuroendocrine phenotypes of prostate cancer via enhancing MET/STAT5A-mediated chromatin accessibility
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits